Chromosome 7HS Research Articles

An island of receptor-like genes at the Rrs13 locus on barley chromosome 6HS co-locate with three novel sources of scald resistance

Three Hordeum spontaneum-derived resistances (referred to as 145L2, 41T1 and 40Y5) have demonstrated long-term effectiveness against barley scald, caused by Rhynchosporium commune, in western Canada. Genetic mapping of these resistances in three populations, and the use of five barley genome assemblies, revealed they co-located to a narrowly defined 0.58–1.2 Mbp region of chromosome 6HS containing the Rrs13 scald resistance gene. Differential disease reactions among the three resistances and a Rrs13 carrier (AB6) to a panel of 24 scald isolates indicated that the four resistances were unique from one another. A marker created to target the 6HS scald locus was screened across a panel of barley germplasm that included H. vulgare, H. spontaneum and H. bulbosum lines. The marker showed specificity to H. vulgare lines known to carry the 6HS scald resistances and to two H. spontaneum lines that trace their origins to Jordan. Within the 0.58–1.2 Mbp region were 2–7 tandemly repeated leucine-rich repeat receptor-like proteins (LRR-RLP) and one lectin receptor-like kinase (Lec-RLK) genes with abundant sequence variation between them. The well-defined role that RLP and RLK genes play in plant defense responses make them logical candidate resistance genes, with one possible hypothesis being that each unique scald resistance may be encoded by a different RLP that interacts with a common RLK. It is suggested the three scald resistances be temporarily named Rrs13145L2, Rrs1341T1 and Rrs1340Y5 to recognize their co-location to the Rrs13 locus until it is determined whether these resistances represent unique genes or alleles of the same gene.

A novel locus conferring resistance to Puccinia hordei maps to the genomic region corresponding to Rph14 on barley chromosome 2HS.

Barley leaf rust (BLR), caused by Puccinia hordei, is best controlled through genetic resistance. An efficient resistance breeding program prioritizes the need to identify, characterize, and map new sources of resistance as well as understanding the effectiveness, structure, and function of resistance genes. In this study, three mapping populations were developed by crossing Israelian barley lines “AGG-396,” “AGG-397,” and “AGG-403” (carrying unknown leaf rust resistance) with a susceptible variety “Gus” to characterize and map resistance. Genetic analysis of phenotypic data from rust testing F3s with a P. hordei pathotype 5457 P+ revealed monogenic inheritance in all three populations. Targeted genotyping-by-sequencing of the three populations detected marker trait associations in the same genomic region on the short arm of chromosome 2H between 39 and 57 Mb (AGG-396/Gus), 44 and 64 Mb (AGG-397/Gus), and 31 and 58 Mb (AGG-403/Gus), suggesting that the resistance in all three lines is likely conferred by the same locus (tentatively designated RphAGG396). Two Kompetitive allele-specific PCR (KASP) markers, HvGBSv2-902 and HvGBSv2-932, defined a genetic distance of 3.8 cM proximal and 7.1 cM distal to RphAGG396, respectively. To increase the marker density at the RphAGG396 locus, 75 CAPS markers were designed between two flanking markers. Integration of marker data resulted in the identification of two critical recombinants and mapping RphAGG396 between markers- Mloc-28 (40.75 Mb) and Mloc-41 (41.92 Mb) narrowing the physical window to 1.17 Mb based on the Morex v2.0 reference genome assembly. To enhance map resolution, 600 F2s were genotyped with markers- Mloc-28 and Mloc-41 and nine recombinants were identified, placing the gene at a genetic distance of 0.5 and 0.2 cM between the two markers, respectively. Two annotated NLR (nucleotide-binding domain leucine-rich repeat) genes (r2.2HG0093020 and r2.2HG0093030) were identified as the best candidates for RphAGG396. A closely linked marker was developed for RphAGG396 that can be used for marker-assisted selection.

High-Resolution Mapping of Barley mild mosaic virus Resistance Gene rym15.

Barley yellow mosaic virus (BaYMV) and Barley mild mosaic virus (BaMMV), which are transmitted by the soil-borne plasmodiophorid Polymyxa graminis, cause high yield losses in barley. In previous studies, the recessive BaMMV resistance gene rym15, derived from the Japanese landrace Chikurin Ibaraki 1, was mapped on chromosome 6HS of Hordeum vulgare. In this study, 423 F4 segmental recombinant inbred lines (RILs) were developed from crosses of Chikurin Ibaraki 1 with two BaMMV-susceptible cultivars, Igri (139 RILs) and Uschi (284 RILs). A set of 32 competitive allele-specific PCR (KASP) assays, designed using single nucleotide polymorphisms (SNPs) from the barley 50 K Illumina Infinium iSelect SNP chip, genotyping by sequencing (GBS) and whole-genome sequencing (WGS), was used as a backbone for construction of two high-resolution maps. Using this approach, the target locus was narrowed down to 0.161 cM and 0.036 cM in the Igri × Chikurin Ibaraki 1 (I × C) and Chikurin Ibaraki 1 × Uschi (C × U) populations, respectively. Corresponding physical intervals of 11.3 Mbp and 0.281 Mbp were calculated for I × C and C × U, respectively, according to the Morex v3 genome sequence. In the 0.281 Mbp target region, six high confidence (HC) and two low confidence (LC) genes were identified. Genome assemblies of BaMMV-susceptible cultivars Igri and Golden Promise from the barley pan-genome, and a HiFi assembly of Chikurin Ibaraki 1 together with re-sequencing data for the six HC and two LC genes in susceptible parental cultivar Uschi revealed functional SNPs between resistant and susceptible genotypes only in two of the HC genes. These SNPs are the most promising candidates for the development of functional markers and the two genes represent promising candidates for functional analysis.

New introgressive forms of cultivated barley obtained on the basis of interspecific hybrids <i>Hordeum vulgare</i> L. × <i>Hordeum bulbosum</i> L.

Background. The use of alien genetic material of bulbous barley Hordeum bulbosum L. to increase the diversity of cultivated barley Hordeum vulgare L. is an important task, since H. bulbosum is characterized by a number of valuable traits. One of the ways to use the genetic potential of bulbous barley is the interspecific hybridization and obtaining fertile introgressive lines of H. vulgare based on interspecific hybrids. The aim of the study was to obtain new introgressive forms of spring barley using interspecific triploid (HvHbHb) and tetraploid (HbHbHvHv) hybrids of cultivated barley with bulbous barley to expand the collection of introgressive lines of H. vulgare. Materials and methods. To create new introgressive forms, diploid barley H. vulgare (2x) cv. ‘Roland’, was crossed with interspecific hybrids H. vulgare cv. ‘Roland’(2x) × H. bulbosum W851 (4х) (HvHbHb), and H. bulbosum A17 (4x) × H. vulgare ‘Borwina’ (4x) (HbHbHvHv). Cultivated barley forms with introgression of the bulbous barley genetic material were selected from the offspring from these crosses; then the selection was continued in two progenies from self-pollination of selected BC1 plants with three alien introgressions. Identification and localization of introgressions was carried out using the method of fluorescent in situ hybridization (GISH and FISH with chromosome-specific markers). Results. The crossing of cultivated barley with triploid and tetraploid interspecific hybrids H. vulgare × H. bulbosum yielded new forms of cultivated barley with recombinant chromosomes, among which two plants with three terminal introgressions of the genetic material of bulbous barley were identified. The first plant, derived from a triploid hybrid, showed introgressions in chromosomes 5HL, 1HL and 3HS. When two homologues with the 5HL introgression of the initial size are combined in the karyotype, the lethality of seedlings is observed in the offspring plants. In the second plant obtained on the basis of a tetraploid hybrid introgression was revealed in chromosomes 5HL, 2HL, and 7HS. In the offspring from self-pollination of this form, the presence of the 2HL introgression of the original size in both homologues led to plant sterility. Forms with a change in size of the introgression in 5HL and 3HS in the offspring of the first plant and with a change in size of the introgression in 2HL in the offspring of the second plant were detected, which indicated that meiotic recombination had occurred in those chromosome regions in BC1 plants. Conclusions A barley plant with the introgression of bulbous barley chromatin into chromosomes 1HL, 5HL, and 3HS of cultivated barley was identified in the offspring from a cross of cultivated barley with a triploid interspecific hybrid H. vulgare × H. bulbosum. In crosses with a tetraploid interspecific hybrid, a barley plant with the introgression into chromosomes 2HL, 5HL, and 7HS was found. On the basis of ‘Roland’ spring barley cultivar, two series of new introgressive forms of H. vulgare with various combinations of recombinant chromosomes have been created.

Delineating the elusive BaMMV resistance gene rym15 in barley by medium-resolution mapping

Barley mild mosaic virus (BaMMV), transmitted by the soil-borne protist Polymyxa graminis, has a serious impact on winter barley production. Previously, the BaMMV resistance gene rym15 was mapped on chromosome 6HS, but the order of flanking markers was non-collinear between different maps. To resolve the position of the flanking markers and to enable map-based cloning of rym15, two medium-resolution mapping populations Igri (susceptible) × Chikurin Ibaraki 1 (resistant) (I × C) and Chikurin Ibaraki 1 × Uschi (susceptible) (C × U), consisting of 342 and 180 F2 plants, respectively, were developed. Efficiency of the mechanical inoculation of susceptible standards varied from 87.5 to 100% and in F2 populations from 90.56 to 93.23%. Phenotyping of F2 plants and corresponding F3 families revealed segregation ratios of 250 s:92r (I × C, χ2 = 0.659) and 140 s:40r (C × U, χ2 = 0.741), suggesting the presence of a single recessive resistance gene. After screening the parents with the 50 K Infinium chip and anchoring corresponding SNPs to the barley reference genome, 8 KASP assays were developed and used to remap the gene. Newly constructed maps revealed a collinear order of markers, thereby allowing the identification of high throughput flanking markers. This study demonstrates how construction of medium-resolution mapping populations in combination with robust phenotyping can efficiently resolve conflicting marker ordering and reduce the size of the target interval. In the reference genome era and genome-wide genotyping era, medium-resolution mapping will help accelerate candidate gene identification for traits where phenotyping is difficult.

Adult resistance genes to barley powdery mildew confer basal penetration resistance associated with broad-spectrum resistance.

Powdery mildew isa major disease of barley (Hordeum vulgare L.) for which breeders have traditionally relied on dominant, pathogen race-specific resistance genes for genetic control. Directional selection pressures in extensive monocultures invariably result in such genes being overcome as the pathogen mutates to evade recognition. This has led to a widespread reliance on fungicides and a single broad-spectrum recessive resistance provided by the mlo gene. The range of resistance genes and alleles found in wild crop relatives and landraces has been reduced in agricultural cultivars through an erosion of genetic diversity during domestication and selective breeding. Three novel major-effect adult plant resistance (APR) genes from landraces, designated Resistance to Blumeria graminis f. sp. hordei (Rbgh1 to Rbgh3), were identified in the terminal regions of barley chromosomes 5HL, 7HS, and 1HS, respectively. The phenotype of the new APR genes showed neither pronounced penetration resistance, nor the spontaneous necrosis and mesophyll cell death typical of mlo resistance, nor a whole epidermal cell hypersensitive response, typical of race-specific resistance. Instead, resistance was localized to the site of attempted penetration in an epidermal cell and was associated with cell wall appositions and cytosolic vesicle-like bodies, and lacked strong induction of reactive oxygen species. The APR genes exhibited differences in vesicle-like body sizes, their distribution, and the extent of localized 3,3-diaminobenzidine staining in individual doubled haploid lines. The results revealed a set of unique basal penetration resistance genes that offer opportunities for combining different resistance mechanisms in breeding programs for robust mildew resistance.

High-resolution mapping of Rym14Hb, a wild relative resistance gene to barley yellow mosaic disease

Key messageWe mapped the Rym14Hb resistance locus to barley yellow mosaic disease in a 2Mbp interval. The co-segregating markers will be instrumental for marker-assisted selection in barley breeding.Barley yellow mosaic disease is caused by Barley yellow mosaic virus and Barley mild mosaic virus and leads to severe yield losses in barley (Hordeum vulgare) in Central Europe and East-Asia. Several resistance loci are used in barley breeding. However, cases of resistance-breaking viral strains are known, raising concerns about the durability of those genes. Rym14Hb is a dominant major resistance gene on chromosome 6HS, originating from barley’s secondary genepool wild relative Hordeum bulbosum. As such, the resistance mechanism may represent a case of non-host resistance, which could enhance its durability. A susceptible barley variety and a resistant H. bulbosum introgression line were crossed to produce a large F2 mapping population (n = 7500), to compensate for a ten-fold reduction in recombination rate compared to intraspecific barley crosses. After high-throughput genotyping, the Rym14Hb locus was assigned to a 2Mbp telomeric interval on chromosome 6HS. The co-segregating markers developed in this study can be used for marker-assisted introgression of this locus into barley elite germplasm with a minimum of linkage drag.

Quantitative trait locus for resistance to the aphid Rhopalosiphum padi L. in barley (Hordeum vulgare L.) is not linked with a genomic region for gramine concentration

Gramine is an indole alkaloid found in certain grass species. Results of previous studies have diverged as to whether this compound might cause resistance to aphids or not. In a breeding program aiming to introduce resistance to the cereal pest Rhopalosiphum padi L. in barley, a cultivar has been crossed with the progenitor of cultivated barley, Hordeum vulgare ssp. spontaneum, and an F1-derived population of doubled haploid (DH) lines was screened both for seedling gramine concentration and resistance to R. padi. The resistance was measured as individual aphid growth in the laboratory. The present study aims to determine if there is a genetic relationship between aphid growth retardation and gramine concentration. To do so, the lines were genotyped with a 384 SNP oligonucleotide pool assay and QTL analyses were performed for both traits. A previously identified aphid resistance locus on the distal part of chromosome 2HS was thereby confirmed, with resistance inherited from H. v. ssp. spontaneum, whereas one or more QTL for gramine concentration were potentially indicated on chromosome 3H, thus corroborating that the two traits are not linked genetically.

Duplicated flavonoid 3'-hydroxylase and flavonoid 3', 5'-hydroxylase genes in barley genome.

BackgroundAnthocyanin compounds playing multiple biological functions can be synthesized in different parts of barley (Hordeum vulgare L.) plant. The diversity of anthocyanin molecules is related with branching the pathway to alternative ways in which dihydroflavonols may be modified either with the help of flavonoid 3′-hydroxylase (F3′H) or flavonoid 3′,5′-hydroxylase (F3′5′H)—the cytochrome P450-dependent monooxygenases. The F3′H and F3′5′H gene families are among the least studied anthocyanin biosynthesis structural genes in barley. The aim of this study was to identify and characterise duplicated copies of the F3′H and F3′5′H genes in the barley genome.ResultsFour copies of the F3′5′H gene (on chromosomes 4HL, 6HL, 6HS and 7HS) and two copies of the F3′H gene (on chromosomes 1HL and 6HS) were identified in barley genome. These copies have either one or two introns. Amino acid sequences analysis demonstrated the presence of the flavonoid hydroxylase-featured conserved motifs in all copies of the F3′H and F3′5′H genes with the exception of F3′5′H-3 carrying a loss-of-function mutation in a conservative cytochrome P450 domain. It was shown that the divergence between F3′H and F3′5′H genes occurred 129 million years ago (MYA) before the emergence of monocot and dicot plant species. The F3′H copy approximately occurred 80 MYA; the appearance of F3′5′H copies occurred 8, 36 and 91 MYA. qRT-PCR analysis revealed the tissue-specific activity for some copies of the studied genes. The F3′H-1 gene was transcribed in aleurone layer, lemma and pericarp (with an increased level in the coloured pericarp), whereas the F3′H-2 gene was expressed in stems only. The F3′5′H-1 gene was expressed only in the aleurone layer, and in a coloured aleurone its expression was 30-fold higher. The transcriptional activity of F3′5′H-2 was detected in different tissues with significantly higher level in uncoloured genotype in contrast to coloured ones. The F3′5′H-3 gene expressed neither in stems nor in aleurone layer, lemma and pericarp. The F3′5′H-4 gene copy was weakly expressed in all tissues analysed.ConclusionF3′H and F3′5′H-coding genes involved in anthocyanin synthesis in H. vulgare were identified and characterised, from which the copies designated F3′H-1, F3′H-2, F3′5′H-1 and F3′5′H-2 demonstrated tissue-specific expression patterns. Information on these modulators of the anthocyanin biosynthesis pathway can be used in future for manipulation with synthesis of diverse anthocyanin compounds in different parts of barley plant. Finding both the copies with tissue-specific expression and a copy undergoing pseudogenization demonstrated rapid evolutionary events tightly related with functional specialization of the duplicated members of the cytochrome P450-dependent monooxygenases gene families.

Forward Genetics Approach Reveals a Mutation in bHLH Transcription Factor-Encoding Gene as the Best Candidate for the Root Hairless Phenotype in Barley.

Root hairs are the part of root architecture contributing significantly to the root surface area. Their role is particularly substantial in maintaining plant growth under stress conditions, however, knowledge on mechanism of root hair differentiation is still limited for majority of crop species, including barley. Here, we report the results of a map-based identification of a candidate gene responsible for the lack of root epidermal cell differentiation, which results in the lack of root hairs in barley. The analysis was based on the root hairless barley mutant rhl1.b, obtained after chemical mutagenesis of spring cultivar ‘Karat’. The rhl1 gene was located in chromosome 7HS in our previous studies. Fine mapping allowed to narrow the interval encompassing rhl1 gene to 3.7 cM, which on physical barley map spans a region of 577 kb. Five high confidence genes are located within this region and their sequencing resulted in the identification of A>T mutation in one candidate, HORVU7Hr1G030250 (MLOC_38567), differing the mutant from its parent variety. The mutation, located in the 3′ splice-junction site, caused the retention of the last intron, 98 bp long, in mRNA of rhl1.b allele. This resulted in the frameshift, the synthesis of 71 abnormal amino acids and introduction of premature STOP codon in mRNA. The mutation was present in the recombinants from the mapping population (F2 rhl1.b × ‘Morex’) that lacked root hairs. The candidate gene encodes a bHLH transcription factor with LRL domain and may be involved in early stages of root hair cell development. We discuss the possible involvement of HORVU7Hr1G030250 in this process, as the best candidate responsible for early stages of rhizodermis differentiation in barley.

A major QTL on chromosome 7HS controls the response of barley seedling to salt stress in the Nure\u2009\xd7\u2009Tremois population

BackgroundSeedling establishment is a crucial and vulnerable stage in the crop life cycle which determines further plant growth. While many studies are available on salt tolerance at the vegetative stage, the mechanisms and genetic bases of salt tolerance during seedling establishment have been poorly investigated. Here, a novel and accurate phenotyping protocol was applied to characterize the response of seedlings to salt stress in two barley cultivars (Nure and Tremois) and their double-haploid population.ResultsThe combined phenotypic data and existing genetic map led to the identification of a new major QTL for root elongation under salt stress on chromosome 7HS, with the parent Nure carrying the favourable allele. Gene-based markers were developed from the rice syntenic genomic region to restrict the QTL interval to Bin2.1 of barley chromosome 7HS. Furthermore, doubled haploid lines with contrasting responses to salt stress revealed different root morphological responses to stress, with the susceptible genotypes exhibiting an overall reduction in root length and volume but an increase in root diameter and root hair density.ConclusionsSalt tolerance at the seedling stage was studied in barley through a comprehensive phenotyping protocol that allowed the detection of a new major QTL on chromosome 7HS.

Genetic dissection of heading date and yield under Mediterranean dry climate in barley (Hordeum vulgare L.)

Understanding the genetics of drought tolerance is a major goal of plant breeders, especially in the Mediterranean region due to its sensitivity to drought and climate changes. Here we investigated genomic regions associated with heading date and yield under rainfed (R) and irrigated (I) conditions, along with some drought tolerance indices derived from phenotypic data collected on a panel of 83 European barley cultivars evaluated in field trials in Foggia, southern Italy, for three growing seasons. A genome wide association analysis (GWAS) was carried out using gene-based SNPs incorporated in the Illumina iSELECT Chip with known map position. A total of 4169 polymorphic SNPs were considered after filtering out SNPs with >10 % missing data and <10 % minimum allele frequency. Heritability was high for heading date and moderate for yield. Principle components analysis partitioned the barley germplasm in three subgroups characterized by different row-type and growth habit. A significant association with heading date was detected on chromosome 2HS, where the different alleles of associated markers were able to significantly affect heading date in different cultivars. Moreover, GWAS uncovered a significant association with yield and relative yield under R conditions on chromosome 5HS. These results provide a clue for further analysis on genetic control of barley flowering time and production in response to water stress under Mediterranean dry climate.

Genetic analysis of grain and malt quality in an elite barley population

Quantitative trait loci (QTLs) associated with grain weight, grain width, kernel hardness and malting quality were mapped in a doubled haploid population derived from two elite Australian malting barley varieties, Navigator and Admiral. A total of 30 QTLs for grain weight, grain width and kernel hardness were identified in three environments, and 63 QTLs were identified for ten malting quality traits in two environments. Three malting quality traits, namely β-amylase, diastatic power and apparent attenuation limit, were mainly controlled by a QTL linked to the Bmy1 gene at the distal end of chromosome 4H encoding a β-amylase enzyme. Six other malting quality traits, namely α-amylase, soluble protein, Kolbach index, free amino-acid nitrogen, wort β-glucan and viscosity, had coincident QTL clustered on chromosomes 1HS, 4HS, 7HS and 7HL, which demonstrated the interdependence of these traits. There was a strong association between these malt quality QTL clusters on chromosomes 1HS and 7HL and the major QTL for kernel hardness, suggesting that the use of this trait to enable early selection for malting quality in breeding programs would be feasible. In contrast, the majority of QTLs for hot-water extract were not coincident with those identified for other malt quality traits, which suggested differences in the mechanism controlling this trait. Novel QTLs have been identified for kernel hardness on chromosomes 2HL and 7HL, hot-water extract on 7HL and wort β-glucan on 6HL, and the resulting markers may be useful for marker-assisted selection in breeding programs.

Barley Ant1 is a homolog of maize C1 and its product is part of the regulatory machinery governing anthocyanin synthesis in the leaf sheath

AbstractAnthocyanins contribute to plants’ defence against a number of abiotic and biotic stress agents. The anthocyanin pigmentation of the barley leaf sheath is genetically determined by Ant1, a gene which maps to a region of chromosome 7HS delimited by the microsatellite loci Xgbms0226 and Xgbms0240. The sequence of the maize gene C1 (encoding an R2R3 MYB factor regulating anthocyanin synthesis) was used for the PCR‐based cloning of Ant1. In ant1 genotypes, no transcript is generated in the leaf sheath, whereas the gene is active in the presence of the dominant allele. A comparison of the coding and promoter sequences of Ant1 (which induces purple pigmentation in the leaf sheath) and ant1 (which does not) showed that the key polymorphisms lay in the promoter sequence. The transcription of four anthocyanin synthesis structural genes (Chi, F3h, Dfr, Ans) was dependent on the allelic status of Ant1.

Rph23: A new designated additive adult plant resistance gene to leaf rust in barley on chromosome 7H

AbstractWe report on a new adult plant resistance (APR) gene Rph23 conferring resistance to leaf rust in barley. The gene was identified and characterized from a doubled haploid population derived from an intercross between the Australian barley varieties Yerong (Y) and Franklin (F). Genetic analysis of adult plant field leaf rust scores of the Y/F population collected over three successive years indicated involvement of two highly additive genes controlling APR, one of which was named Rph23. The gene was mapped to chromosome 7HS positioned at a genetic distance 36.6 cM. Rph23 is closely linked to marker Ebmac0603, which is flanked by markers bPb‐8660 and bPb‐9601 with linkage distances of 0.8 and 5.1 cM, respectively. A PCR‐based marker was optimized for marker‐assisted selection of Rph23, and on the basis of this marker, the gene was postulated as being common in Australian and global barley germplasm. Pedigree and molecular marker analyses indicated that the six‐rowed black Russian landrace ‘LV‐Taganrog’ is the likely origin of Rph23.

The genetic relationship between barley leaf rust resistance genes located on chromosome 2HS

Four seedling leaf rust resistance genes (Rph14, Rph15, Rph16 and Rph17) described in barley are located on chromosome 2HS. A fifth, present in barley accession HOR 1063, was reported to map to the same region as Rph16. A comprehensive analysis of the allelic relationship between Rph14, Rph15, Rph17 and RphHOR1063 was undertaken. Two additional genotypes (HOR 15560 and Zhu 4) that were initially thought to carry Rph16 were also included. While no segregation was observed in F2 populations derived from crosses between PI 584760 (carrying Rph14), PI 355447 (carrying Rph15), Zhu 4 and HOR 1063, the lines did exhibit differential reactions across 10 Australian Puccinia hordei pathotypes. This suggests that the Rph14 locus may be complex. An evaluation of F3 populations confirmed that resistance genes in Zhu 4 and in HOR 1063 were allelic to both Rph14 and Rph15, however A susceptible line was found among F3 progeny of the cross PI 584760/PI 355447, suggesting that Rph14 and Rph15 may be independent. Rph17 and the gene present in HOR 15560 were shown to be independent of each other and of Rph14 and Rph15. The response of HOR 15560 across an array of P. hordei pathotypes revealed that it possesses the same specificity as Rph1. Because the resistance in Zhu 4 and HOR 1063 were shown to be allelic to Rph14, they should be designated Rph14.am and Rph14.an, respectively.

Towards positional isolation of three quantitative trait loci conferring resistance to powdery mildew in two Spanish barley landraces.

Three quantitative trait loci (QTL) conferring broad spectrum resistance to powdery mildew, caused by the fungus Blumeria graminis f. sp. hordei, were previously identified on chromosomes 7HS, 7HL and 6HL in the Spanish barley landrace-derived lines SBCC097 and SBCC145. In the present work, a genome-wide putative linear gene index of barley (Genome Zipper) and the first draft of the physical, genetic and functional sequence of the barley genome were used to go one step further in the shortening and explicit demarcation on the barley genome of these regions conferring resistance to powdery mildew as well as in the identification of candidate genes. First, a comparative analysis of the target regions to the barley Genome Zippers of chromosomes 7H and 6H allowed the development of 25 new gene-based molecular markers, which slightly better delimit the QTL intervals. These new markers provided the framework for anchoring of genetic and physical maps, figuring out the outline of the barley genome at the target regions in SBCC097 and SBCC145. The outermost flanking markers of QTLs on 7HS, 7HL and 6HL defined a physical area of 4 Mb, 3.7 Mb and 3.2 Mb, respectively. In total, 21, 10 and 16 genes on 7HS, 7HL and 6HL, respectively, could be interpreted as potential candidates to explain the resistance to powdery mildew, as they encode proteins of related functions with respect to the known pathogen defense-related processes. The majority of these were annotated as belonging to the NBS-LRR class or protein kinase family.

Fine mapping and comparative genomics integration of two quantitative trait loci controlling resistance to powdery mildew in a Spanish barley landrace

The intervals containing two major quantitative trait loci (QTL) from a Spanish barley landrace conferring broad spectrum resistance to Blumeria graminis were subjected to marker saturation. First, all the available information on recently developed marker resources for barley was exploited. Then, a comparative genomic analysis of the QTL regions with other sequenced grass model species was performed. As a result of the first step, 32 new markers were added to the previous map and new flanking markers closer to both QTL were identified. Next, syntenic integration revealed that the barley target regions showed homology with regions on chromosome 6 of rice (Oryza sativa), chromosome 10 of Sorghum bicolor and chromosome 1 of Brachypodium distachyon. A nested insertion of ancestral syntenic blocks on Brachypodium chromosome 1 was confirmed. Based on sequence information of the most likely candidate orthologous genes, 23 new barley unigene-derived markers were developed and mapped within the barley target regions. The assessment of colinearity revealed an inversion on chromosome 7HL of barley compared to the other three grass species, and nearly perfect colinearity on chromosome 7HS. This two-step marker enrichment allowed for the refinement of the two QTL into much smaller intervals. Inspection of all predicted proteins for the barley unigenes identified within the QTL intervals did not reveal the presence of resistance gene candidates. This study demonstrates the usefulness of sequenced genomes for fine mapping and paves the way for the use of these two loci in barley breeding programs.

Genetic mapping and QTL analysis of disease resistance traits in the barley population Baudin×AC Metcalfe

A genetic map of barley (Hordeum vulgare L.) with 163 amplified fragment length polymorphism and 69 simple sequence repeat (SSR) markers was constructed using a population of 178 doubled haploid lines from a cross between the varieties ‘Baudin’ and ‘AC Metcalfe’. Linkage groups were assigned to barley chromosomes using published map locations of SSR markers as reference points. The total length of the map was 1307.2 cM with an average interval length of 5.6 cM between markers. The genetic map was used to locate quantitative trait loci (QTLs) for several disease resistance traits. The population was tested for spot type net blotch, caused by Pyrenophora teres f. maculata, and net type net blotch, caused by Pyrenophora teres f. teres, in greenhouse experiments and in a range of field environments in Western Australia and Queensland. The response of the lines to leaf rust (caused by Puccinia hordei L.) at adult plant growth stages was evaluated in Western Australia. Significant marker–trait associations were found for seedling resistance and symptom severity in adult plants to these diseases. A new locus conferring resistance to P. teres f. maculata at both seedling and adult plant stages was detected on the short arm of chromosome 6H. From the seedling testing against P. teres f. teres, five highly repeatable QTLs were detected, on chromosomes 2HS, 2HL, 3HS, 4HL, and 6HS. For leaf rust, one highly significant QTL was detected on chromosome 2HL. The markers within these QTL regions present an opportunity for marker-assisted selection for these traits in barley-breeding programs.

Detection of photoperiod responsive and non-responsive flowering time QTL in barley

A QTL analysis was performed to determine the inheritance of flowering time in barley, using a set of recombinant inbred lines developed from a winter-type × spring-type cross. Two photoperiod responsive loci, Ppd-H1 and Ppd-H2, were detected on chromosome arms 2HS and 1HL respectively. Segregation for eam8 (mapping to the terminus of chromosome arm 1HL) and Eam5 (close to Sgh2 on chromosome arm 5HL) was also observed. These latter two genes functioned under 12 h to 24 h photoperiods. In addition, eps2S and eps7S, known to lie on chromosome arms 2HS and 7HS respectively, were detected. A new QTL for flowering time, qDHE.ak-1HS, was mapped 23 cM from the terminus of chromosome 1HS, and appears to be expressed under extremely short day lengths.